“DNA FINGERPRINTING IN HUMAN HEALTH AND

SOCIETY ”

BY
RAVINDRA KUMAR PANCHAL
(08BT000122)
&
AGRAWAL PARTH TARAPRAKASH
(08BT000105)

GUIDED BY:
Dr. NAVNEET SINGH CHAUDHARY

Submitted in partial fulfillment of the Degree of Bachelor of Technology

SESSION 2010-2011

DEPARTMENT OF BIO TECNOLOGY

SIR PADAMPA T SINGHANIA UNIVERSITY UDAIPUR

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ACKNOWLEDGMENT
First and foremost, we would like to express my sincere gratitude to my report guide, Dr.
Navneet Singh Chaudhary . We were privileged to experience a sustained enthusiastic and
involved interest from his side. This fuelled our enthusiasm even further and encouraged us
to boldly step into what was a totally dark and unexplored expanse before us.
We would also like to thank our seniors who were ready with a posi-tive comment all the
time, whether it was an off-hand comment to encour-age us or a constructive piece of
criticism.
We would like to thank the SPSU staff members and the institute, in general, for extending a
helping hand at every juncture of need. We would like to express our gratitude towards our
parents for their kind co-operation and encouragement which help us in completion of this
project.
Last but not least, our thanks and appreciations also go to our col-leagues in developing the
project and people who have willingly helped us out with their abilities.

CONTENTS
1. Introduction………………………………………………………… 4-10
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1.1 Abstract 4
1.2 Executive summary 4-5
1.3 Report objective 5-6
1.4 dna(deoxyribonucleicacid) 7
1.5 about dna fingerprinting 8
1.6 what is dna fingerprinting 9
1.7 principles of dna fingerprinting10
1.7.1 base pairing10
2. methods of dna fingerprinting ……………………….…………….11-34
2.1 RFLP 12-27
2.1.1 steps of RFLP 14-26
 Isolation of DNA.15
 Cutting, sizing, and sorting. 15-22
 Transfer of DNA to nylon.22-24
 Probing.25
2.1.2 analysis of dna fingerprints26-27

2.2 PCR
2.3 AmpFLP
2.4.STR
3. applications of dna fingerprinting………………………………....35-43
3.1 dna forensic.
3.2 Paternity Testing.
3.3 MIA Soldiers.
3.4 Inherited Disorder Testing.
3.5 Personal Identification.
3.6 DNA Forensics Databases .

4. Conclusion……………………………………………………………….44
5. References………………………………………………………………………………………….45

CHAPTER 1:INTRODUCTION
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1.1 ABSTRACT
This project investigated the impact of the new technology of DNA fingerprinting on society,
especially the legal system and database ethics. Our conclusions propose an expansion of
DNA databases to include individuals convicted of any felony, not just violent crimes. The
benefit to society of such DNA databases to identify unknown corpses, determine paternity,
place a suspect at the scene of a crime, develop leads where otherwise there were none, and
link crimes to identify serial criminals outweighs any privacy issues especially for convicted
felons.

1.2 EXECUTIVE SUMMARY

Five decades ago, Watson and Crick discovered the secrets of DNA structure. DNA
Fingerprinting, or DNA profiling, was first adopted in 1984 by Oxford University educated
Alec Jeffreys. Jeffrey's discovery opened a whole new world that, once proven and perfected,
would unlock markers to visualize each person's unique identity. Inside each human being, as
well as plants, animals, and microorganisms, lies a unique DNA structure. Today, DNA
Fingerprinting is "rapidly becoming the primary method for identifying and distinguishing
individual human beings” (Betsch, 1994). Some of the applications of DNA fingerprinting
techniques include: murder cases, rape cases, paternity testing, diagnosis of inherited
disorders, military identification, and molecular archaeology. Forensic DNA analysis has
been admitted into United States courtrooms since 1987. DNA profiling does not claim to be
an absolute identification, but may be very strong evidence and is considered to be just one
part of the entire case. DNA profiling is used primarily in sexual assault cases. Prior to DNA
testing, labs were limited regarding the amount of genetic information that could be obtained
from evidence samples. Now with DNA testing, the genetic content of the evidence sample
itself can be examined. In the case of a rape investigation, four profiles are formed: the first is
a DNA profile from the blood of the victim, the second DNA profile is formed from the
blood of the defendant, thirdly a vaginal swab is taken, and the female and male fractions are
separated and profiled separately. The victim’s blood profile and the vaginal swab profile
should match alleles. The defendant’s blood sample profile should match the male fraction
profile in order to consider him as a suspect. It is important to remember that DNA profiling
does not incriminate a suspect with absolute certainty but if the profiles don’t 5 match it can

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exclude with absolute certainty. In the case of likely matches, statistics are used to determine
the frequency that this DNA profile likely appears in the general human population
.
"Since the discovery of DNA fingerprinting at the turn of the 20th century, science has
assumed an increasingly important and powerful role in the decision making process of our
judicial branch" (Biancamano, 1996). Many Landmark cases, which set the standards for
admitting DNA into the courtroom, as well as the reliability and acceptability of DNA
techniques, will be discussed to prove the methods used today are valid and reliable when
performed properly. Some of the earlier cases that will be talked about did not actually
involve DNA, but what they did accomplish was to set precedence by establishing new rules
for admittance of technical evidence, and to set new generally accepted scientific standards in
the court of law. Some of these landmark cases include: Frye v. United States in 1923,
Federal Rules of Evidence 702 (Rule 702) in 1975, Colin Pitchfork in 1986, Andrews v
Florida in 1988, People v Castro in 1989, Two Bulls v US in 1990, and the case of Paul
Eugene Robinson in 2003 where the DNA evidence was the primary basis of the conviction
of sexual assault.

1.3 REPORT OBJECTIVE:

DNA fingerprinting is a powerful new technology, which is used to assist in convicting

the guilty and exonerating the innocent. The topic of DNA fingerprinting however remains
controversial in the courtroom regarding technical issues, and also has legal, cultural and
political consequences. This controversy is mainly derived from the lack of knowledge
regarding the procedures of DNA fingerprinting, how it is obtained, analyzed, and reported.
The purpose of this project is to help to eliminate the public’s doubt, concerning DNA
fingerprinting and help them to convert their reservations about the science into support for
the use of DNA fingerprinting in the courtroom. The focus of this paper will be directed
toward the layperson in order to target the prospective population of jurors. Jurors are
members of our community who are required to evaluate the relevance of all evidence
including DNA evidence. Thus, once they are on a jury, and DNA evidence is brought in,
they will be able to comprehend the data and make an informed verdict. This paper will cover
the primary areas of the public’s concerns, as well as provide history of DNA analysis, and
future advances of the science for admission into the courtroom. Description of the lab report,
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as well as the actual data used to formulate the lab report will help the public develop an
appreciation for the formulation of the results of the actual testing. The objective of this paper
was accomplished by first, outlining the science of the main techniques of DNA
fingerprinting in laymen’s terms. Secondly the project describes the processes of collection,
storage, and prevention of contamination of the DNA evidence. Thirdly, landmark DNA
court cases that established legal precedents for admitting DNA in U.S. courts will be
revisited and analyzed. Lastly, many of the controversies regarding the DNA databases will
be analyzed. This project not only informs the reader about the facts of the history of DNA 8
fingerprinting, but it will also entice the reader to encourage and support the use of DNA
evidence in the courtroom.The overall goal of this paper is for the reader to walk away with
an understanding of DNA fingerprinting’s positive impact on society.

1.4 DNA(Deoxyribonucleic acid):

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Deoxyribonucleic acid, more commonly known as DNA, is the complex chemical structure
that uniquely identifies each and every organism. An organism’s complete set of DNA is
known as a genome. DNA is the fundamental building block of the genome (An Introduction
to DNA, 2002). DNA is located inside an organism's chromosomes.

"DNA itself is a double-helix polymer. A polymer is simply a large molecule that is

produced by several smaller units linking together and forming a chain" (Rohloff, 2000).

Figure – Watson and Crick’sdiscovery of the DNA “DoubleHelix” (Devitt, 2002)

1.5 ABOUT DNA FINGERPRINTING

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DNA Fingerprinting, or DNA profiling, was first adopted in 1984 by Oxford University
educated Alec Jeffreys. Jeffreys stumbled upon this theory while working on myoglobin, the
gene that codes for an iron-containing protein in muscles.
Jeffrey's discovery opened a whole new world that, once proven and perfected, would unlock
each person's unique identity. Inside each and every human being, as well as plants, animals,
and microorganisms, lies a unique DNA structure .Conventional fingerprints occur only on
the fingertips, and are capable of being altered, but a DNA fingerprint is the same for"every
cell, tissue, and organ of a person”. DNA is incapable of being altered, and is "rapidly
becoming the primary method for identifying and distinguishing among individual human
beings”. In science, DNA Fingerprinting does not point to a unique individual. However, it
provides a profile, and then the probability that there are others who also match the profile is
determined leading to the match or conviction. So what is DNA fingerprinting used for?
Some of the applications of DNA fingerprinting techniques include: murder cases, rape cases,
paternity testing, diagnosis of inherited disorders, military identification, and molecular
archaeology.
In this chapter, we will explore each application, as well as simplify DNA for the average
person, show how to run a fingerprint, trace its growth and impact on society, and look
toward future uses such as national DNA databases and DNA fingerprints as one day
becoming our National Identification Cards.

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Fig:-The clues to murder mysteries and crime are found in the minute evidence of blood
and body fluids. The solution to many crimes is found in DNA 'fingerprinting'

1.6 WHAT ARE DNA FINGERPRINTS?

DNA profiling (also called DNA testing, DNA typing, or genetic fingerprinting) is a
technique employed by forensic scientists to assist in the identification of individuals by their
respective DNA profiles. DNA profiles are encrypted sets of numbers that reflect a person's
DNA makeup, which can also be used as the person's identifier.

Although 99.9% of human DNA sequences are the same in every person, enough of the DNA
is different to distinguish one individual from another. DNA profiling uses repetitive
("repeat") sequences that are highly variable, called variable number tandem repeats (VNTR).
VNTRs loci are very similar between closely related humans, but so variable that unrelated
individuals are extremely unlikely to have the same VNTRs.

1.7 Principles of DNA fingerprinting:

1.7.1 Base pairing;-

Each type of base on one strand forms a bond with just one type of base on the other strand.
This is called complementary base pairing. Here, purines form hydrogen bonds to
pyrimidines, with A bonding only to T, and C bonding only to G. This arrangement of two
nucleotides binding together across the double helix is called a base pair. As hydrogen bonds
are not covalent, they can be broken and rejoined relatively easily. The two strands of DNA
in a double helix can therefore be pulled apart like a zipper, either by a mechanical force or
high temperature. As a result of this complementarity, all the information in the double-
stranded sequence of a DNA helix is duplicated on each strand, which is vital in DNA
replication. Indeed, this reversible and specific interaction between complementary base pairs
is critical for all the functions of DNA in living organisms.

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Top, a GC base pair with three hydrogen bonds. Bottom, an AT base pair with two hydrogen
bonds. Non-covalent hydrogen bonds between the pairs are shown as dashed lines.

The two types of base pairs form different numbers of hydrogen bonds, AT forming two
hydrogen bonds, and GC forming three hydrogen bonds (see figures, left). DNA with high
GC-content is more stable than DNA with low GC-content, but contrary to popular belief,
this is not due to the extra hydrogen bond of a GC base pair but rather the contribution of
stacking interactions (hydrogen bonding merely provides specificity of the pairing, not
stability).

CHAPTER2:“METHODS OF DNA FINGERPRINTING”

RFLP:
Restriction fragment length polymorphism (RFLP) analyzes the length of the strands of the
DNA molecules with repeating base pair patterns. DNA molecules are long strands found
tightly wound in chromosomes which are contained in the nucleus of each human cell.
Within each DNA strand are numbers of genes that determine the particular characteristics of
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an individual. While about 5% of the gene compositions on DNA contain this type of genetic
information, the other 95% do not. However, of the 95%, these non-coding genes contain
identifiable repetitive sequences of base pairs, which are called Variable Number Tandem
Repeats (VNTR). To extract a DNA fingerprint, a Southern blot is performed and the DNA
is analyzed via a radioactive probe. The restriction fragment length polymorphism analysis is
used to detect the repeated sequences by determining a specific pattern to the VNTR, which
becomes the person's DNA fingerprint. The drawback with this system is that it requires a
considerable amount of DNA in order to be used.

PCR
The polymerase chain reaction (PCR) was developed by Karry Mullis of the Cetus
Corporation in 1983 for use in research laboratories for establishing hereditary
authentication. The PCR analysis amplifies the DNA molecules using a smaller sample. On
the forensic front, the PCR found to be useful in identifying DNA fingerprints in criminal
matters and in paternity tests because it requires less amounts of DNA because it makes
identical copies of the DNA sample. The PCR analysis amplified isolated regions on the
strands of the DNA under examination. The drawback was that it was not as discriminating
as the RFLP.

AmpFLP
Amplified fragment length polymorphism (AmpFLP) came into vogue in the 90's and is still
popular in the smaller countries involved in the process of DNA fingerprinting. It remains
attractive because of its relatively less complicated operation and the cost-effectiveness of
the procedure. By using the PCR analysis to amplify the minisatellite loci of the human cell,
this method proved quicker in recovery than the RFLP. However, due to the use of gel in its
analysis phase, there are issues of bunching of the VTRN's, causing misidentifications in the
process.

STR
The short tandem repeat (STR) methodology for extracting DNA is the system most widely
used form of DNA fingerprinting. This system is based on the features of PCR, as it utilizes
specific areas that have short sequential repeat DNA. The STR analyzes how many times
base pairs repeat themselves on a particular location on a strand of DNA. The big advantage

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in this method is that the DNA comparisons can match the possibilities into an almost
endlessrange.

DNA fingerprinting has been extremely successful for use in the personal identification of
criminal suspects, paternity issues, as well as in identification of the deceased. DNA,
however, still poses issues because the VNTRs are not evenly distributed in all people
because they are inherited. In addition, there is still the human element as the final voice in
the administration of all DNA fingerprinting procedures. However, as forensic science
continues its work in DNA testing, there appears to be no limit to the value it can render to
society.

2.1Process of Making a DNA Profile using RFLP method:-

The key to DNA profiling is to make a comparison of unique loci of the DNA left at the
crime scene with a suspect’s DNA. The portion of the genome where there is a lot of
diversity among individuals is called polymorphic regions. The polymorphic regions used for
forensics are the non-coding regions. These are the regions of the DNA that do not code for
proteins and they make-up 95% of our genetic DNA. These regions are therefore called the
“junk” portion of the DNA. Although these “junk” regions do not generate proteins, they can
regulate gene expression, they aid in the reading of other genes that do formulate the proteins,
and they are a large portion of the chromosome structure (How DNA Evidence Works, 2004).
The non-coding DNA regions are made-up of length polymorphisms, which are variations in
the physical length of the DNA molecule. The DNA profile analyzes the length
polymorphisms in the non-coding areas. These polymorphisms are identical repeat sequences
of DNA base pairs. The number of tandem repeats at specific loci on the chromosome varies
between individuals. For any specific loci, there will be a certain number of repeats. These
repeat regions are classified into groups depending on the size of the repeat region. Variable
number of tandem repeats (VNTRs) have repeats of 9-80 base pairs. Short tandem repeats
(STRs) contain 2-5 base pair repeats (Brief Introduction to STRs, 2004). For each of our 23
pairs of chromosomes, we inherit one copy of each chromosome from our mother and the
other from our father. “This means that you have two copies of each VNTR locus, just like
you have two copies of real genes (Figure-4). If you have the same number of sequence
repeats at a particular VNTR site, you are called homozygous at that site; if you have a

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different number of repeats, you are said to be heterozygous” (How DNA Evidence Works,
2001).

Figure 4: The diagram above displays three possible VNTR combinations.

DNA analysis is a laboratory procedure that requires a number of steps. There are a number
of techniques used by different laboratories, however in this paper two techniques will be
reviewed: Restriction Fragment Length Polymorphism (RFLP), and Polymerase Chain
Reaction (PCR) using Short Tandem Repeats (STRs).
The first step in both procedures involves the extraction and purification of the DNA. Before
a DNA sample can be analyzed, the DNA needs to be isolated from the other organic and
non-organic portions of the sample. The type of sample will determine the technique used to
isolate the DNA. The sample may be boiled with a detergent that breaks down the proteins
and other cellular material but does not affect the DNA. Enzymes may be added to break
down proteins and other cellular material. Organic solvents may be used to separate the DNA
from the other organic and non-organic material. The DNA is then separated from the
proteins and othercellular material (DNA Forensics, Problem Set 1, 2004).

2.1.1Steps for DNA fingerprinting by RFLP method

1. Isolation of DNA.
2. Cutting, sizing, and sorting.
3. Transfer of DNA to nylon.
4. Probing.
5. DNA fingerprint.

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 Fig : showing the basic steps of the dna fingerprinting

Step 1: Isolation of DNA:-

The process begins with a sample of an individual's DNA (typically called a "reference
sample"). The most desirable method of collecting a reference sample is the use of a buccal
swab, as this reduces the possibility of contamination. When this is not available (e.g.
because a court order may be needed and not obtainable) other methods may need to be used
to collect a sample of blood, saliva, semen, or other appropriate fluid or tissue from personal
items (e.g. toothbrush, razor, etc.) or from stored samples (e.g. banked sperm or biopsy
tissue). Samples obtained from blood relatives (biological relative) can provide an indication
of an individual's profile, as could human remains which had been previously profiled.

 DNA can be extracted from almost any human tissue.

 Buccal cells from inside cheek for paternity tests.
 Sources of DNA at crime scene: blood, semen, hair follicle, saliva.
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  DNA extracted from evidence is compared to DNA from known individuals.

Step2:Cutting, sizing, and sorting

Restriction enzymes are enzymes isolated from bacteria that recognize specific sequences in
DNA and then cut the DNA to produce fragments, called restriction fragments. Restriction
enzymes play a very important role in the construction of recombinant DNA molecules, as is
done in gene cloning experiments. Another application of restriction enzymes is to map the
locations of restriction sites in DNA. Special enzymes called restriction enzymes are used to
cut the DNA at specific places. For example, an enzyme called EcoR1, found in bacteria, will
cut DNA only when the sequence GAATTC occurs. The DNA pieces are sorted according to
size by a sieving technique called electrophoresis. The DNA pieces are passed through a gel
made from seaweed agarose (a jelly-like product made from seaweed). This technique is the
DNA equivalent of screening sand through progressively finer mesh screens to determine
particle sizes.

Fig:- showing the digestion of dna fragment by the restriction enzymes

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Sepration of dna fragments by GEL ELECTROPHORESIS
according to their size:

Agarose gel electrophoresis is a method used in biochemistry and molecular biology to

separate a mixed population of DNA and RNA fragments by length, to estimate the size of
DNA and RNA fragments or to separate proteins by charge. Nucleic acid molecules are
separated by applying an electric field to move the negatively charged molecules through an
agarose matrix. Shorter molecules move faster and migrate farther than longer ones because
shorter molecules migrate more easily through the pores of the gel. This phenomenon is
called sieving. Proteins are separated by charge in agarose because the pores of the gel are
too large to sieve proteins.

"Electrophoresis" refers to the electromotive force (EMF) that is used to move the molecules
through the gel matrix. By placing the molecules in wells in the gel and applying an electric
field, the molecules will move through the matrix at different rates, determined largely by
their mass when the charge to mass ratio (Z) of all species is uniform, toward the anode if
negatively charged or toward the cathode if positively charged.

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17
Materials required for agarose gel electrophorosis

Typically 10-30 μl/sample of the DNA fragments to separate are obtained, as well as a
mixture of DNA fragments (usually 10-20) of known size (after processing with DNA size
markers either from a commercial source or prepared manually).

 Buffer solution, usually TBE buffer or TAE 1.0x, pH 8.0 .

 Agarose .
 An ultraviolet-fluorescent dye, ethidium bromide, (5.25 mg/ml in H2O). The stock
solution be careful handling this. Alternative dyes may be used, such as SYBR Green.
 Nitrile rubber gloves. Latex gloves do not protect well from ethidium bromide .
 A color marker dye containing a low molecular weight dye such as "bromophenol
blue" (to enable tracking the progress of the electrophoresis) and glycerol (to make
the DNA solution denser so it will sink into the wells of the gel).
 A gel rack.
 A "comb" .
 Power Supply .
 UV lamp or UV lightbox or other method to visualize DNA in the gel .

Preparation

There are several methods for preparing gels. A common example is shown here. Other
methods might differ in the buffering system used, the sample size to be loaded, the total
volume of the gel (typically thickness is kept to a constant amount while length and breadth
are varied as needed). Most agarose gels used in modern biochemistry and molecular biology
are prepared and run horizontally.

1. Make a 1% agarose solution in 100ml TAE, for typical DNA fragments (see figures).
A solution of up to 2-4% can be used if you analyze small DNA molecules, and for
large molecules, a solution as low as 0.7% can be used.
2. Carefully bring the solution just to the boil to dissolve the agarose, preferably in a
microwave oven.

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 3. Let the solution cool down to about 60 °C at room temperature, or water bath. Stir or
swirl the solution while cooling.

Wear gloves from here on, ethidium bromide is a mutagen, for more information on safety
see ethidium bromide

1. Add 5 µl ethidium bromide (from a stock solution of 10 mg/ml) per 100 ml gel
solution for a final concentration of 0.5 µg/ml. Be very careful when handling the
concentrated stock. Some researchers prefer not to add ethidium bromide to the gel
itself, instead soaking the gel in an ethidium bromide solution after running, in this
case the ethidium bromide solution has final concentration of 0.5 µg/ml.
2. Stir the solution to disperse the ethidium bromide, then pour it into the gel rack.
3. Insert the comb at one side of the gel, about 5–10 mm from the end of the gel.
4. When the gel has cooled down and become solid, carefully remove the comb. The
holes that remain in the gel are the wells or slots.
5. Put the gel, together with the rack, into a tank with TAE. Ethidium bromide at the
same concentration can be added to the buffer. The gel must be completely covered
with TAE, with the slots at the end electrode that will have the negative current.

Procedure

After the gel has been prepared, use a micropipette to inject about 2.5 µl of stained DNA (a
DNA ladder is also highly recommended). Close the lid of the electrophoresis chamber and
apply current (typically 100 V for 30 minutes with 15 ml of gel). The colored dye in the DNA
ladder and DNA samples acts as a "front wave" that runs faster than the DNA itself. When
the "front wave" approaches the end of the gel, the current is stopped. The DNA is stained
with ethidium bromide, and is then visible under ultraviolet light.

1. The agarose gel with three slots/wells (S).

2. Injection of DNA ladder (molecular weight markers) into the first slot.
3. DNA ladder injected. Injection of samples into the second and third slot.
4. A current is applied. The DNA moves toward the positive anode due to the negative
charges on its phosphate backbone.
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5. Small DNA strands move fast, large DNA strands move slowly through the gel. The
DNA is not normally visible during this process, so the marker dye is added to the
DNA to avoid the DNA being run entirely off the gel. The marker dye has a low
molecular weight, and migrates faster than the DNA, so as long as the marker has not
run past the end of the gel, the DNA will still be in the gel.
6. Add the color marker dye to the DNA ladder

After the electrophoresis is complete, the molecules in the gel can be stained to make
them visible. Ethidium bromide, silver, or Coomassie Brilliant Blue dye may be used for
this process. Other methods may also be used to visualize the separation of the mixture's
components on the gel. If the analyte molecules fluoresce under ultraviolet light, a
photograph can be taken of the gel under ultraviolet lighting conditions, often using a Gel
Doc. If the molecules to be separated contain radioactivity added for visibility, an
autoradiogram can be recorded of the gel.

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 Fig:- Gel electrophoresis result

Step 3; Transfer of dna fragment from gel to nitrocellulose

membrane:-

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A Southern blot is a method routinely used in molecular biology for detection of a
specific DNA sequence in DNA samples.

The original method was first developed by E. M. Southern in 1975 and later was modified
by various people for various applications. The process of Southern blotting starts after the
agarose gel electrophoresis of restriction digest or DNA. The gel is first washed with buffer
to remove the broken or fragmented residues of agarose formed during banding and the
accumulated contaminants.

Then the gel is placed in acid solution (0.25 M HCI) for about 5-10 min. During this step, the
DNA undergoes depurination. After this step the gel is placed in 0.25 M NaOH alkali
solution. This step ensures that the DNA is shorter in length (<500 bp) and is in single-
stranded form. DNA fragments large in size (>10 kb) require a longer time and move in zig-
zag form when compared to shorter DNA fragments.

Finally, the gel is extensively washed with buffer to remove the traces of HCI and NaOH.
Then the gel is placed on the filter paper with wigs dipped in a reservoir containing transfer
buffer placed in transfer buffer reservoir tank which is placed on the glass plate.

Nitrocellulose or nylon filter paper is placed on the gel above which a stack of blotting
papers soaked in transfer buffer is placed and gently pressed using glass plate or rod to
remove the air that is trapped between the gel and membrane. Then a stack of unsoaked
blotting papers is placed above them and the glass plate with a weight of 500 g is placed
above it.

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By capillary action the transfer buffer is transferred from the reservoir to the blotting papers.
During this process the DNA from the gel is transferred onto the nitrocellulose paper and
immobilized there due to the negative charge of the DNA and the positive charge of the
membrane.

This set up is allowed to stand like that for 12-18 hours. After this time period the
nitrocellulose or nylon membrane is taken out from the set up carefully. As the binding of the
DNA to the nitrocellulose or nylon membrane is weak, it has to be fixed more strongly, so
that the DNA is not washed off during the washing of the membranes. There are two widely
used methods for this procedure.
After the transfer, the membrane is exposed to ultraviolet rays, so that a bond is formed

between thymine residues present in the DNA and the positively charged amino groups on
the surface of the nylon membrane. This method is called as UVcross linking method. The
other method is baking of the nitrocellulose or nylon membrane at 80°C in a vacuum oven.

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Step 4: Probing

Adding radioactive or colored probes to the nylon sheet produces a pattern called the DNA
fingerprint. Each probe typically sticks in only one or two specific places on the nylon sheet.

In molecular biology, a hybridization probe is a fragment of DNA or RNA of variable

length (usually 100-1000 bases long), which is used in DNA or RNA samples to detect the
presence of nucleotide sequences (the DNA target) that are complementary to the sequence in
the probe. The probe there by hybridizes to single-stranded nucleic acid (DNA or RNA)
whose base sequence allows probe-target base pairing due to complementarity between the
probe and target. The labeled probe is first denatured (by heating or under alkaline conditions
such as exposure to sodium hydroxide) into single DNA strands and then hybridized to the
target DNA (Southern blotting) or RNA (northern blotting) immobilized on a membrane or in
situ.

Let's start with the mechanism, how the probe identifies the sequence. Normally DNA is in
double stranded form, and the nucleotides A-T and G-C will form 2 and 3 hydrogen bonds to
each other (A to T and C to G). When we heat the DNA, it will denaturate to single stranded
form and hydrogen bonds will no more bond the strands together, so we will have two
complementary strands.

Now, lets add a short piece of single stranded DNA which is labeled with radioactive
phosphorus or sulfur*. If the probe's sequence is ATTCCGG...TT, it will slide past DNA
which doesn't have complementary sequence. When our probe happens to hit the sequence
AA...CCGGAAT, it will attach firmly to its "missing half". Of course most probe molecules
will never find their complementary sequences and will attach to TT...CCCGAAT, for
example. But we can adjust the environment so that those probes that are not firmly attached
will break off, and only the true sequences are found. This separation can be done by
adjusting temperature and salt consentration etc., collectively referred to as the "stringency".

In Southern blotting we have first separated genomic DNA from a tissue sample. Then the
long genomic DNA is cut into smaller parts with restriction enzymes. Pieces are separated
according to their size with agarose gel electrophoresis and then transferred to a membrane,
usually nylon or nitrocellulose.

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Now that we have our DNA bound to a solid surface, separated by size, we denaturate the
DNA and add the radiolabeled probe. The probe attaches to it's counterpart, some to almost
their counterpart, and some just wander over the membrane. Then we just wash away the
probes which are loosely attached to DNA, because the more complementary the sequence is,
the more hydrogen bonds can form and the stronger the "union" that will form between these
two molecules. And after washing we just expose the membrane to x-ray film and can see, if
we, for example, were succesfull to achieve a transgenic calf.

2.1.2ANALYSIS OF DNA FINGERPRINTS:

DNA fingerprinting is often used in criminal cases to determine a suspect's guilt. DNA from
a blood, hair root, or semen sample found at the scene of a crime can prove guilt or innocence
with high precision. The DNA sample is cleaved with a restriction enzyme and the resulting
fragments are separated using Southern blotting techniques. DNA from the crime scene is
analyzed on the same gel as DNA from the potential criminals, and therefore, DNA collected
from the scene can be compared with that from various suspects and the RFLP's produced
from the DNA of the guilty suspect will match with the RFLP's produced from DNA
collected from the crime scene.

In the following example, a woman was raped while she and her fiance were sleeping in their
car. They were found the next morning in the woods next to a recreation area and both had
died of gunwounds. One man was later found driving the stolen vehicle and he told
authorities of the friend that was with him the night of the murders. In order to determine
which suspect was guilty of raping the woman, DNA fingerprinting (RFLP analysis) was
used. DNA from a semen sample retrieved from the body was compared with DNA from
blood samples from both suspects. These DNA samples were cut with a particular restriction
enzyme and fragments were separated by gel electrophoresis. Figure 3 shows the RFLP's
from the semen sample in yellow and the RFLP's from both suspects in purple and blue. It is
shown in this figure that suspect 2 is the man that raped this woman. It is possible that an
innocent person could show the exact same restriction fragments, but the chance of this is 1
in 9,390,000,000, which is twice the human population of the world! Suspect 2 was conviced
of rape and murder and received a double death sentence. This happened to be the first case
in the world in which the conviction of the death sentence was based on DNA fingerprinting
(The Dolan DNA Learning Center, 2000).

25
 

Fig:- RFLP analysis of the semen sample collected from the raped woman versus blood
samples of two suspects. The pink and blue fragments were produced from genomic DNA
from the suspects and the yellow fragments were produced from genomic DNA from the
victim. This figure was reproduced pending permission by the

2.2“ AFLP-PCR:Amplified Fragment Length Polymorphism ”

26
Amplified Fragment Length Polymorphism PCR (or AFLP-PCR or just AFLP) is a
PCR-based tool used in genetics research, DNA fingerprinting, and in the practice of genetic
engineering. Developed in the early 1990s by Keygene, AFLP uses restriction enzymes to
digest genomic DNA, followed by ligation of adaptors to the sticky ends of the restriction
fragments. A subset of the restriction fragments is then selected to be amplified. This
selection is achieved by using primers complementary to the adaptor sequence, the restriction
site sequence and a few nucleotides inside the restriction site fragments (as described in detail
below). The amplified fragments are visualized on denaturing polyacrylamide gels either
through autoradiography or fluorescence methodologies.

AFLP-PCR is a highly sensitive method for detecting polymorphisms in DNA. The technique
was originally described by Vos and Zabeau in 1993. In detail, the procedure of this
technique is divided into three steps:

1. Digestion of total cellular DNA with one or more restriction enzymes and ligation of
restriction half-site specific adaptors to all restriction fragments.
2. Selective amplification of some of these fragments with two PCR primers that have
corresponding adaptor and restriction site specific sequences.
3. Electrophoretic separation of amplicons on a gel matrix, followed by visualisation of
the band pattern.

A variation on AFLP is cDNA-AFLP, which is used to quantify differences in gene

expression levels.

Another variation on AFLP is TE Display, used to detect transposable element mobility.

27
 FIG:-Example of AFLP Data from a Capillary Electrophoresis Instrument

2.3 Polymerase Chain Reaction (PCR)

PCR is a new technique that was developed in 1986 by Kary Mullis and was applied to
forensics in 1991 (Forensic Fact Files DNA Profiling, 2004). PCR has the ability to replicate
genetic material, and even proofread and make corrections on the copies (Forensic Fact Files
DNA Profiling, 2004). It involves the repeated copying of specified areas of DNA molecules.
These areas are the alleles and are specific sequences of base pairs. These target areas are the
variable regions. At either end of these target areas are “flanker” bases that are the non
variable regions. The flankers occur at the same locations on the chromosomes of all people.
The basis of PCR is to reproduce thousands of copies of a particular variable region of the
DNA that is of interest. This results in DNA fragments that vary in length due to the
variability in the number of repeated regions. The process is used to amplify a small amount
of DNA taken from a sample of blood, hair, etc. With each cycle of copying, the quantity of
the target allele is doubled resulting in an exponential amplification of the gene in the PCR.
After 30 copying cycles, there will be one billion times more copies of the target alleles than
at the start of the amplification process (Principle of the PCR, 1999).

28
Figure : Steps in PCR. (Principle of PCR, 2004

29
There are three steps in the PCR process (see Figure):

1) Denaturation: This process takes place at 94 degrees Celcius. This stage separates the
DNA double helix, and forms DNA single strands. This also forces every enzymatic reaction
to come to a halt.

2) Annealing: This usually takes place at 54 degrees Celsius. The primers bind to their
complementary bases on the now single-stranded DNA.

3) Extension: This takes place at 72 degrees Celsius. This step involves the synthesis of
DNA by polymerase. Starting from the primer, the polymerase reads the template strand and
matches it with complementary bases. The result is two new helixes, each composed of one
of the original strands plus its newly assembled complementary strand (Principle of the PCR,
1999).

In order to obtain more copies of DNA, the three step process is repeated. Each phase only
takes 1-3 minutes, thus if you completed the process for 45 minutes, millions of copies could
be generated because every time the process is repeated the amount of DNA doubles
(exponentially) (Figure-8).

FIG: Diagram of Exponential Growth in the Number of DNA Copies During PCR.

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2.3.1DNAFingerprintingAnalysis
This DAF analysis system is used routinely to characterise unknown isolates of Foc from
Australia and worldwide by comparing the DNA fingerprints of these unknown isolates with
those from our DNA reference collection (which spans all the currently identified VCGs of
Foc). This DAF analysis system has been thoroughly optimised and provides an informative
and reproducible platform for the molecular characterisation of unknown isolates of Foc.

Genome-specific DNA fingerprint patterns are analysed and scored manually (by eye);
fingerprints of test samples are compared to standards (characterised isolates) by
assessing the general fingerprint pattern and scoring for presence/absence of polymorphic
bands (see Figure 8). Scoring is made easier by placing gels on a lightbox, which enables the
discrimination of lightly stained bands. Test samples can be compared with standards from
other gels by overlapping gels on the lightbox, or by digitally cutting and pasting the lanes
next to one to build a new image that makes scoring easier. DAF analysis is used in
conjunction with VCG testing for the characterisation of unknown isolates of Foc.

Figure 8. DNA Fingerprinting analysis of a range of Australian and overseas VCGs of Foc,
visualised by polyacrylamide gel electrophoresis and silver staining. The DAF PCR was
performed using the HIRH single oligonucleotide primer. This DAF analysis clearly shows
the utility of this technique for differentiating closely related VCGs of Foc. Isolate
31
information is annotated on the gel photograph. Image courtesy of the CRC for Tropical
Plant Protection.

Analysis of the alleles uses length variation of the amplified alleles. The alleles used are
repeated units of the same sequence of bases. The length variation is the number of times that
a particular base sequence is repeated between flankers. PCR techniques are divided into
different categories depending on the number of base pairs in the repeated units. Variable
Number of Tandem Repeats (VNTR) uses between 9 and 40 bases per unit; Amplified
Fragment Length Polymorphism (AmpFLP) uses between 8 and 16 bases per unit; and Short
Tandem Repeats (STR) uses between 4 and 6 bases per unit (Schumm, 1996). STRs will be
focused on in this paper because it is the technique of choice for most forensics laboratories
presently.

2.4.“STRs(short tenden repeats)”

The short tandem repeat (STR) methodology for extracting DNA is the system most widely
used form of DNA fingerprinting. This system is based on the features of PCR, as it utilizes
specific areas that have short sequential repeat DNA. The STR analyzes how many times
base pairs repeat themselves on a particular location on a strand of DNA. The big advantage
in this method is that the DNA comparisons can match the possibilities into an almost
endlessrange.

During forensic examination, an STR with a known repeat sequence is extracted and
separated by electrophoresis. The distance that the STR migrated is examined. However,
capillary columns are now used rather than the gel electrophoresis that was used in the RFLP
analysis. Two short pieces of synthetic DNA called primers are specially designed to attach
to a conserved common non-variable region of DNA, which flanks the variable target region
of the DNA. A mixture of chemicals including the primers, the individual bases and a
copying enzyme (polymerase) is added to the solution with the original DNA strand that is to
be copied. The primers are short chains of the bases that make up the single strand of DNA.
Primers are complementary to the conserved regions flanking the targeted STR (black arrows
in Figure-6). The primer used in the PCR amplification process attaches a fluorescent tag to
the target alleles enabling the distance traveled by the DNA fragments during capillary
32
electrophoresis to be detected using a fluorescent scanner (Blackett Family DNA Activity 2,
2004).

CHAPTER 3: “APPLICATIONS OF DNA

FINGERPRINTING”

33
3.1 DNA FORENSICS:

Forensic DNA analysis has been admitted into United States courtrooms since 1987. The
common question being answered is “Who is the source of this biological material?” (Inman
and Rudin, 1997). DNA profiling was first used in the United States courtroom in the case of
Tommie Lee Andrews. (He was suspected of serial rape, therefore, the investigators wanted
to link his DNA to the DNA of the 23 victims. Because of the DNA analysis technology and
the CODIS system, Andrews was linked to the series of rapes, and sentenced to 115 years
(Ramsland, 2004). Profiling has helped to acquit or convict suspects of rape and/or murder
however it is used in less than one percent of all criminal cases (Genetic Science Learning
Center, 2004). DNA profiling does not claim to be an absolute identification, but may be very
strong evidence and is considered to be just one part of the entire case. DNA profiling is used
primarily in sexual assault cases. Prior to DNA testing, labs were limited regarding the
34
amount of genetic information that could be obtained from semen. Now with DNA testing,
the genetic content of the sperm itself can be examined. The RFLP method of analysis needs
a considerable amount of material for testing and may be unable to determine anything from
a sample that has any degradation. The PCR analysis needs only a minute amount of sample
because it replicates the DNA and can work even with degraded samples (Sullivan Personal
Interview 2004). In the case of a rape investigation four profiles are formed (Figure-10): the
first is a DNA profile from the blood of the victim, the second DNA profile is formed from
the blood of the defendant, thirdly a vaginal swab is taken, and the female and male fractions
are separated and profiled separately. The victim’s blood profile and the vaginal swab profile
should match alleles. The defendant’s blood sample profile should match the male fraction
profile in order to consider him as a suspect. It is important to remember that DNA profiling
does not incriminate a suspect it can only exclude suspects.

Some examples of DNA uses for forensic identification include:

• Identify potential suspects whose DNA may match evidence left at crime scenes
• Exonerate persons wrongly accused of crimes
• Identify crime and catastrophe victims
• Establish paternity and other family relationships
• Identify endangered and protected species as an aid to wildlife officials (could be used for
prosecuting poachers)

35
• Detect bacteria and other organisms that may pollute air, water, soil and food
• Match organ donors with recipients in transplant programs
• Determine pedigree for see or livestock breeds
• Authenticate consumables such as caviar and wine
(The FBI’s DNA Databasing Initiatives, 2000)

Murder or Rape Applications

DNA Fingerprinting has evolved to take part in a plethora of different applications. Thefirst,
and most well known application is forensics, either murder or rape cases. Suppose anofficer
arrived at a crime scene of a murder, and the only evidence that could be found was a
bloodstain on the carpet from the suspect. If you were put on a jury and told, based on
scientific evidence using DNA Fingerprinting, that this suspect was the murderer, would you
convict him? In the beginning stages of its existence, courts would not always allow the
evidence to be admitted, based on the fact it hadn't been completely proven to be accurate. As
you'll see in a following chapter, many court cases and laws continued to set precedence, and
DNA Fingerprinting eventually became more and more accessible and less controversial in
the court of law. The FBI began to establish DNA Fingerprinting as means of connection to
crimes early on. Between the years of 1989 and 1996, they used the technique in about
10,000 sexual assault cases (Establishing Innocence, 2004). In those cases, 2,000 prime
suspects were proven innocent based on the results of the DNA profiling (Establishing
Innocence, 2004). In Figure 8, you can see an example of an RFLP fingerprint analysis of 7
suspects relative to a crime scene bloodstain. It seems very clear that Suspect 3 was present at
the crime scene. It is also true to say that without this method of testing, those 2,000 innocent
prime suspects could most likely have faced conviction (Establishing Innocence, 2004).
Many cases from decades ago have been recently reopened and their verdicts changed by
admitting new DNA evidence. Many of these cases will be explained in depth in a later
chapter.

36
Figure 20 – Evidence that is now used thanks to DNA Fingerprinting techniques.(Huskey, 1995)

3.2 Paternity Testing

Paternity testing has also become a very commonly used application of DNA Fingerprinting.
When a child is born, the child inherits 23 chromosomes from the mother, and chromosomes
from the father. Therefore, when a DNA test is done, “the visible band pattern of the child is
unique. Half matches the mother and half matches the father”. A DNA paternity test is the
most accurate form of testing possible to determine parentage. If the patterns do not match on
two or more probes, then the father is 100% excluded, which means he is without question
not the father. If a match is present on every DNA probe, the probability of paternity is 99.9%
or greater. In a Court of Law, 99% is accepted as proof of paternity (DNA Diagnostic Center,
2004). In Figure 9, you can see that the mother (blue bands) and father (orange bands) both
have separate DNA patterns. Now look at daughter 1. You can see that some of the mothers
same blue bands are inherited as well as some of the fathers orange bands. Daughter 2 has the
mothers blue bands, but not the orange band, instead 38 red bands. This proves that the father
of daughter 1 is not the father of daughter 2, but a father from the mother’s previous
marriage. Son 1 is a child of both of the shown parents as well. Son 2, however, is adopted
37
because he doesn't have any of the parents DNA. One of the most famous paternity cases
involved Thomas Jefferson, the third President of the United States, who in 1802, was
accused of impregnating Sally Hemings, a slave at the Jefferson estate. No verification or
conviction was ever brought about however. The story was sustained throughout the decades,
and in 1998, Dr. Eugene Foster and a team of geneticists, conducted DNA tests to prove the
accuracy of these accusations. The test results proved that it was indeed a Jefferson who
fathered Sally Heming's son, Eston. At the time of the child’s birth, there where
approximately twenty-five Jefferson's living in Virginia, all of who carried the
chromosome that would match the child's. The verdict proved that it was probable, yet not
conclusive, that Thomas Jefferson was the father of Eston Hemings (The Plantation, 2004).

Figure 21 – A DNA
Paternity Test showing
results after being
tested using DNA
Fingerprints. (Antler,
2004)

3.3 MIA Soldiers:

Another application of DNA Fingerprinting is for historical clarifications and identifications.

Over the last decade, hundreds of missing in action (MIA) soldiers, that were never
identified, have been identified through the use of DNA Fingerprinting. How is this possible?
Relatives of unidentified soldiers who are willing to use a sample kit to submit a blood and
38
saliva sample to the laboratory, where “forensic anthropologists, forensic dentists and
equipment recovery specialists who work at the U.S. Army Central Identification
Laboratory” (Tippy, 2004) analyze the sample, and are able to match DNA with MIAs, with
as little as a bone fragment recovered from a killed soldier (Tippy, 2004).
"The most famous American memorial for unidentified soldiers killed in combat is located at
Arlington National Cemetery in Arlington, Virginia" (Unknown Soldier). It has inherited the
name as the Tomb of the Unknown Soldier (Unknown Soldier, 2004). In 1998, the remains of
soldiers killed during the Vietnam War, which occurred between 1959 and 1975, were
removed for hope of identification. After testing was completed, the remains were identified
as First Lieutenant Michael Blassie, an Air Force pilot who was killed when his helicopter
was shot down in Vietnam in 1972. After the positive identification, his family buried the
remains in Saint Louis, near Blassie's childhood home. In 1999, Pentagon officials made the
decision that based on technological advances in this DNA profiling and the ability to
identify MIA's, no other soldier from the Vietnam War would be buried in the Tomb of the
Unknown Soldier (Unknown Soldier, 2004).

3.4 Inherited Disorder Testing

DNA Fingerprinting is also capable of determining inherited disorders in adults, children, and
unborn babies. A historical example to prove this involves our sixteenth president and the
issue of Marfan's Syndrome (Betsch, 1994), a disease which is characterized by unusually
long and nimble limbs. As you know, President Lincoln was abnormally tall during his
lifetime which lasted from 1809 to 1865. Could DNA evidence from Lincoln be recovered
that would prove this disease existed? According to David F. Betsch, "the technology is so
powerful that even the blood-stained clothing from Abraham Lincoln has been analyzed for
evidence of a genetic disorder called Marfan's Syndrome" (Betsch, 1994). Although many
attempts have been made to clearly identify this disorder in Abraham Lincoln, no conclusive
result has been reached due to damaging of historical artifacts. This genetic testing allows
people to "test for some 40 anomaly that flags a disease or disorder” (Inherited Disorders,
2002). “Much of the current excitement in gene testing, however, centers on predictive gene
testing: tests that identify people who are at risk of getting a disease, before any symptoms
appear” (Inherited Disorders, 2002).

39
3.5 Personal Identification

DNA fingerprinting has seen tremendous growth over the last couple of decades, but is there
more on the horizon for new applications? One of the main challenges in the field is to create
an ideal system of personal identification that would be “indelible, unalterable and –unlike an
ID card –part of the individual” (Marx, 1998). The scientific goal is to generate this as a
national standard, where every citizen is included within the database, and to prove that there
are no drawbacks, such as misuse of medical information during the identification process, or
any cautious steps that are taken before DNA fingerprinting becomes the standard (Marx,
1998). In 2002, Alec Jeffreys, the inventor of DNA fingerprinting, put forth his theory on the
DNA databases. He believed that “every citizen’s genetic information should be stored on the
UK national register” (O'Brien, 2002). Jeffreys went on to say that “if we’re all on the
database, we’re all in exactly the same boat – the issue of discrimination disappears”
(O'Brien, 2002). The advantages and disadvantages of DNA databases, as well as its progress
in the modern day society, will be explained in more depth in a following chapter. Another
challenge in this growing field is to simplify and hasten the procedure of running a DNA
fingerprint which will revolutionize forensics. In Australia, researchers have claimed that
they have a technique that is twice as fast as conventional DNA technology and is capable of
cranking out 1,000 samples a day, where most laboratories manage only a few dozen
(Dayton, 2002). “Moreover, profiles are derived from DNA contained in a single cell —
existing 41 procedures require at least 500 cells” (Dayton, 2002). Team leader, Ian Findlay,
of the Australian Genome Research Facility, said of his team’s recent discoveries, that “it’s
such a breakthrough we’re waiting for the rest of the world to catch up.” (Dayton, 2002) So
this is where the debate begins. In the chapters to follow, forensics of DNA, landmark DNA
court cases, and DNA databases will be examined. Also issues such as the validity of these
tests, and the use within the courtroom and why some courts still will not completely trust
DNA evidence in the courtroom.

3.6 DNA Forensics Databases

40
National,DNADatabank:CODIS:

The COmbined DNA Index System, CODIS, blends computer and DNA technologies into a
tool for fighting violent crime. The current version of CODIS uses two indexes to generate
investigative leads in crimes where biological evidence is recovered from the crime scene.
The Convicted Offender index contains DNA profiles of individuals convicted of felony sex
offenses (and other violent crimes). The Forensic index contains DNA profiles developed
from crime scene evidence. All DNA profiles stored in CODIS are generated using STR
(short tandem repeat) analysis.

CODIS utilizes computer software to automatically search its two indexes for matching DNA
profiles. Law enforcement agencies at federal, state, and local levels take DNA from
biological evidence (e.g., blood and saliva) gathered in crimes that have no suspect and
compare it to the DNA in the profiles stored in the CODIS systems. If a match is made
between a sample and a stored profile, CODIS can identify the perpetrator.

This technology is authorized by the DNA Identification Act of 1994. All 50 states have laws
requiring that DNA profiles of certain offenders be sent to CODIS. As of January 2003, the
database contained more than a million DNA profiles in its Convicted Offender Index and
about 48,000 DNA profiles collected from crime scenes but which have not been connected
to a particular offender.

As more offender DNA samples are collected and law enforcement becomes better trained
and equipped to collect DNA samples at crime scenes, the backlog of samples awaitning
testing throughout the criminal justice system has increased dramatically. In March 2003
President Bush proposed $1 billion in funding over 5 years to reduce the DNA testing
backlog, build crime lab capacity, stimulate research and development, support training,
protect the innocent, and identify missing persons. For more information, seethe U.S.
Department of Justice's.

Potential Benefits of DNA Databanking Arrestee:

41
  Most major crimes involve people who also have committed minor offenses.
 Innocent people currently are incarcerated for crimes they did not commit; if samples
had been taken at the time of arrest, these individuals would have been excluded early
in the investigative process.
 Moving the point of testing from conviction to arrest would result in savings in
investigation, prosecution, and incarceration.
 Investigators would be able to compare other cases against the arrested person's DNA
profile, just as with fingerprints.

CHAPTER 4: CONCLUSION

This report has been an effort to bring to the public a greater understanding of DNA
fingerprinting and its uses. To begin our examination we looked at DNA structure and the

42
process of creating a DNA profile. To continue with the paper we explored the stringent steps
that must be taken to collect and preserve forensic evidence to help its acceptance in the
courtroom. It logically followed that we should inspect the historical significance of the
precedence set by landmark court cases concerning the admissibility of technical evidence
including DNA profiles. Finally we looked towards the future with the establishment and
ongoing legislative modifications of the CODIS system. We chose these subjects to focus on
because it is important to inform the general public of them in order to dispel any myths
regarding the usage of DNA in forensics. We felt that this topic provided a perfect example
of a new technology with great impact on society, worthy of an IQP. The advances in
forensic technology have allowed the sophistication and standardization of the process by
which forensic evidence is collected and preserved in order to increase the chances of its
acceptance in the courtroom. We have found that due to the nature of most forensic evidence,
heat and moisture can cause severe degradation to samples. Because of this it is necessary to
avoid packaging DNA evidence in plastic containers because plastic does not allow moisture
to escape, and the DNA degrades. Also, because contamination, especially in the case of PCR
analysis can render the resulting profile virtually useless, it is essential that only clean
instruments are used in the collection, and that care is taken to collect elimination control
samples. Chain of custody must also be established to avoid any argument of tampering. The
importance of the treatment of this genetic material cannot be overstressed as the world
painfully 77 learned in the OJ Simpson trial. Without the rigid guidelines that are now in
place, it would be difficult to argue the credibility of the evidence itself.

43